Involvement of ERK, p38 and NF-kappaB signal transduction in regulation of TLR2, TLR4 and TLR9 gene expression induced by lipopolysaccharide in mouse dendritic cells

Toll-like receptors (TLR) are sentinel receptors capable of recognizing pathogen-associated molecule patterns (PAMP) such as lipopolysaccharide (LPS) and CpG-containing oligonucleotides (CpG ODN). TLR2 and TLR4 are major receptors for Gram-positive and Gram-negative bacterial cell wall components, respectively. TLR9 is necessary for CpG signalling. LPS or CpG ODN can activate immature dendritic cells (DC) and induce DC maturation characterized by production of cytokines, up-regulation of co-stimulatory molecules, and increased ability to activate T cells. However, little is known regarding the regulation of TLR gene expression in mouse DC. In this study, we investigated the regulation of TLR2, TLR4 and TLR9 gene expression by LPS in murine immature DC. TLR2, TLR4 and TLR9 mRNA were up-regulated following LPS stimulation. The up-regulation of TLR9 expression coincided with significantly increased production of tumour necrosis factor-alpha induced by LPS plus CpG ODN. While inhibition of extracellular signal-related kinase and NF-kappaB activation suppressed the up-regulation of the expression of TLR2, TLR4 and TLR9 mRNA, inhibition of p38 kinase prevented the up-regulation of TLR2 and TLR4 mRNA expression but enhanced the up-regulation of TLR9 expression. These results demonstrated that TLR2, TLR4 and TLR9 gene expression was differently regulated by LPS in mouse immature DC. Up-regulation of TLR2, TLR4 and TLR9 expression by LPS might promote the overall responses of DC to bacteria and help to explain the synergy between LPS and other bacterial products in the induction of cytokine production.     

小儿肝的年龄解剖学研究

分六个年龄组观察了180个(男98,女82)小儿肝的形态、度量、位置及体表投影.小儿肝相对较大,右叶大于左叶,下界位置较低并逐渐上移.在右锁骨中线的肋弓下一般均可触到.小儿肝重随其年龄(身高)的增长而增长,其增长速度相对较慢,尤以Ⅶ组小儿更明显.     

Screening of human anti-TNF-alpha scFv from large phage library

AIM: To clone human anti-TNF-alpha scFv from large phage antibody library. METHODS: Panning of large phage library against TNF-alpha was conducted to select specific antibodies. The antigen binding activity and DNA sequence were determined and analyzed by ELISA, restriction enzyme map. RESULTS: After 4 rounds of panning, 62 positive clones were obtained and 27 clones had specific binding ability to TNF-alpha. DNA fingerprinting of the 27 clones showed 7 different stripes indicated 7 different positive clones. 5 of the 7 clones expressed soluble anti-TNF-alpha activity. DNA sequence analysis showed that the variable regions of these scFvs belonged to different subgroups. CONCLUSION: Human TNF-alpha scFv have been successfully obtained from large phage antibody library.     

Translocation of Enterococcus faecalis strains across a monolayer of polarized human enterocyte-like T84 cells

We used a two-chamber system to study transcytosis of Enterococcus faecalis across monolayers of human colon carcinoma-derived T84 cells, which show structural resemblance to the native intestine. Among 16 E. faecalis isolates from different sources, the well-characterized strain OG1RF and 8 other isolates (2 endocarditis isolates, 1 urine isolate, and all 5 fecal isolates) showed translocation in this assay, while 6 clinical isolates (3 endocarditis and 3 urine isolates), the recipient strain JH2-2, and the control, Escherichia coli DH5alpha, had no detectable translocation. Of two OG1RF mutants involving the previously studied epa (enterococcal polysaccharide antigen) gene cluster, known to be needed for virulence and resistance to killing by polymorphonuclear leukocytes, one epa mutant (TX5179) was unable to translocate, while TX5180, with an epa disruption farther downstream, showed a moderate decrease in translocation relative to that of the wild-type strain OG1RF (P < 0.01), indicating that the epa gene cluster is important for translocation across a T84 monolayer. This observation was confirmed by complementation of the epa mutant (TX5179) with epa genes and restoration of its translocation ability. In conclusion, we have demonstrated translocation of at least some strains of E. faecalis across T84 monolayers, although strains differ considerably in this ability, and we have demonstrated that epa mutations can cause marked changes in successful translocation. These results suggest that this model may be a useful in vitro system for studying the process of translocation from the intestinal tract.     

涎腺癌肉瘤临床及病理分析

目的：探讨涎腺癌肉瘤的临床病理学特点及其鉴别诊断。方法：对3例涎腺癌肉瘤患者的临床资料进行回顾性研究并复习相关文献，对全部病例的组织学标本重新进行镜下观察。结果：涎腺癌肉瘤临床表现常为迅速增大的颜面部肿物并伴疼豢。光匀下组织学观察常可见肉瘤和癌两种成分并存，癌多为鳞状细胞癌和腺癌，肉瘤以骨或软骨肉瘤为主。结论：涎腺癌肉瘤的临床特点与涎腺其他恶性肿瘤较难区别，但涎腺癌肉瘤的恶性程度极高。     

奥美拉唑对家兔肾脏内髓集合管细胞H+/K+交换功能的影响

目的探讨奥美拉唑对家兔肾脏IMCD细胞H+/K+交换的影响,以及经洗涤处理是否解除这种影响.方法原代培养家兔肾脏IMCD细胞单层,在100 μmol/L奥美拉唑缓冲液中孵育25 min, 洗涤组则在奥美拉唑缓冲液孵育后,用不含奥美拉唑的缓冲液洗涤IMCD细胞;对照组在不含奥美拉唑的缓冲液孵育25 min;CECF/AM荧光探针法测定各组IMCD细胞H+/K+交换.结果奥美拉唑组的H+/K+交换为(0.016±0.006) dpHi/min(n=8);与对照组的(0.053±0.008) dpHi/min(n=8)相比,相差非常显著(P<0.001);洗涤组的H+/K+交换为(0.016±0.006) dpHi/min(n=6),与对照组(n=6)的(0.052±0.009)dpHi/min相比,相差非常显著(P<0.001).结论 100 mol/L的奥美拉唑对家兔IMCD细胞的H+/K+交换有显著影响,而且洗涤处理不能解除这种抑制.因而,肺心病合并上消化道出血患者使用奥美拉唑时,应综合考虑其利弊.     

Identification of a new HLA-B*56 variant, B*5614

In this report, the novel allele B*5614 is presented. The allele was identified in a Chinese individual by sequence-based typing. HLA-B*5614 differs from B*5608 by a single nucleotide at position 277G-->C in exon 2. This results in an amino acid change from Gly to Arg at codon 93.     

Application of Serum-Free Culture Medium for Preparation of A-NK Cells

To compare the differences between proliferation and cytotoxicity of adherent natural killer （A-NK） cells cultured with serum-free medium AIMV and standard serum-containing medium in vitro, and also observe the assisting effect of IL-12 on the activation and the morphology character of IL-2-treated A-NK cells, cellular proliferation was evaluated by MTT method in vitro. The morphology of the target cells killed by A-NK cells was observed through electroscope. All of the A-NK cells cultured in serum-free medium AIMV could rapidly proliferate and keep high cytotoxicity compared with that in standard serum-containing medium. A-NK cells activated by both moderate-dose IL-2 and IL-12 were superior to the high-dose IL-2-treated A-NK cells. These data indicated that serum-free medium AIMV could replace standard serum-containing medium for culturing A-NK cells, and moderate-dose IL-2 and IL-12 could reduce side effects caused by high-dose IL-2. The study provided a new experimental basis for experimental and clinical preparation of A-NK cells. Cellular ＆ Molecular Immunology.